Solid phase synthesis supports and methods

ABSTRACT

Functionalized supports and methods for solid phase synthesis. Preferably, the functionalized support is azlactone-functionalized.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. Ser. No. 09/827,107, filed Apr.5, 2001, now U.S. Pat. No. 6,787,635, the disclosure of which is hereinincorporated by reference.

BACKGROUND

The recent surge of interest in combinatorial chemistry and automatedsynthesis has created a renewed interest in polymer-supported reactions.Combinatorial chemistry is a synthetic strategy that leads to largechemical libraries by the systematic and repetitive covalent connectionof a set of different “building blocks” of varying structures to eachother to yield a large “library” of diverse molecules. It isparticularly useful in producing polypeptides or polynucleotides thatare currently of interest in the biotechnology area. Polymer-supportedreactions or solid phase synthesis is the main methodology used incombinatorial chemistry.

In order to perform combinatorial chemistry in the solid phase, thestarting materials are covalently bonded to a polymeric support.Reagents can then be added that react with the starting materials toyield products that are still attached to the support. The mainadvantage of solid phase synthesis is that the products don't need to bepurified. They can be retained on the solid phase while excess reagentsand byproducts are washed away. Then, by successive treatment withdifferent reagents, new molecules are built up on the solid phase. Byusing a variety of starting materials it is possible to simultaneouslybuild up a library of related compounds by using a single reagent or setof reagents. In this way many new products can be produced in a singlereaction vessel.

A wide variety of materials have been developed as polymeric supportsand are commercially available. Most of these materials are based onlightly crosslinked polystyrene, a relatively hydrophobic polymer. Thecrosslinker most commonly used has been divinylbenzene. Crosslinkingimproves the mechanical properties of the resin but prevents swelling ofthe resin, which is essential for rapid and thorough reactivity withinthe polymer system. The hydrophobicity of polystyrene limits itsusefulness in many solvents and with many reagents. In order to overcomethe problems associated with hydrophobicity, a more hydrophilicmaterial, such as polyethylene glycol (PEG) has been coated onto orgrafted to the polystyrene to make it more versatile for solid phasesynthesis. This is a very expensive process and still does notcompletely address the problems associated with the hydrophobicpolystyrene matrix. Polystyrene resins have also been crosslinked withmore hydrophilic crosslinkers such as bifunctional styrene derivatizedPEG chains to crosslink polystyrene in order to improve general resinperformance. Improved swelling and mechanical properties have beenobserved with these resins. However, PEG-based crosslinkers cannot beused with strong bases or organometallic reagents; thus, theirusefulness is limited. Additionally, many prior art matrices used forsolid phase synthesis have generally low crosslink density and aregel-type polymers. This polymer structure, however, can lead to problemsrelated to reagent diffusion during synthesis. Thus, new and improvedresins that can be used for solid phase synthesis are needed.

The purpose of combinatorial chemistry is to generate a large library ofrelated compounds in order to test them for a desired property. Forinstance, in the drug industry, there is an interest in screening alarge number of related compounds for biological activity. Usually thesecompounds are screened after cleavage from the support. Under thesecircumstances, the synthesis of combinatorial libraries requiresimmobilization of the first building block to the support via a linkerand cleavage of the compound from the linker after the library synthesisis complete.

The linker is a molecule that can be permanently attached to the supportvia covalent bonds and also has a reactive group capable of binding, forexample, the first building block molecule of the intended syntheticlibrary. After the first building block is attached, further groups aresystematically added sequentially until a terminal building block isattached. Finally, the desired library molecules are cleaved from thelinker and thus the support. Chloromethylated crosslinked polystyrene isconventionally used to immobilize carboxylic acid building blocks via anunsubstituted benzyl ester. However, these unsubstituted benzyl-typelinkers require harsh cleavage conditions, usually liquid HF. There is aneed for new linker-functionalized supports that are stable to thereaction conditions used to build the library molecules on the support,but are also able to form an easily cleavable bond with the librarymolecule under mild conditions to release those compounds at the end ofthe synthesis.

SUMMARY OF THE INVENTION

The present invention provides functionalized supports and methods foruse for solid phase synthesis, which are useful in combinatorialchemistry, for example. Functionalized supports described herein can bein the form of a plurality of particles or a membrane, for example.Furthermore, the functionalized support can form a combinatorial libraryin preferred embodiments.

Generally, preferred functionalized support material (with linkerincorporated therein, herein referred to as a “linker-functionalizedsupport”) of the present invention has the formulaSS—[NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m) wherein SS represents asupport material; R¹, R², R³, and R⁴ are each independently hydrogen oran organic group (preferably, a (C1–C14)alkyl group, a(C3–C14)cycloalkyl group, or a (C5–C12)aryl group) with the proviso thatat least one of R³ and R⁴ is an aromatic group (preferably, a(C5–C12)aryl group); R⁷ is hydrogen or an organic group (preferably,including a reactive site, which may optionally be protected by aprotecting group); p is at least 1 (preferably, 1 to 20, and morepreferably, 1 to 2); and m is 1 to the resin capacity of the supportmaterial. Typically and preferably, theNH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) groups are bound to the supportmaterial through a carbonyl group.

Alternatively, preferred functionalized support material (with linkerincorporated therein) of the present invention has the formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein SS represents a support material; R¹, R², R³, and R⁴ are eachindependently hydrogen or an organic group (preferably, a (C1–C14)alkylgroup, a (C3–C14)cycloalkyl group, or a (C5–C12)aryl group) with theproviso that at least one of R³ and R⁴ is an aromatic group (preferably,a (C5–C12)aryl group); R⁵ and R⁶ are each independently an organic group(preferably, a (C1–C14)alkyl group, a (C3–C14)cycloalkyl group, or a(C5–C12)aryl group); R⁷ is hydrogen or an organic group (preferably,including a reactive site, which may optionally be protected by aprotecting group); n is 0 to 1; p is at least 1 (preferably, 1 to 20,and more preferably, 1 to 2); and m is 1 to the resin capacity of thesupport material. This material is a preferred example of anazlactone-functionalized support material having a linker attachedthereto, wherein C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from anazlactone group.

Yet another preferred functionalized support material (with linkerincorporated therein) of the present invention has the formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(R⁸)—NH—C(O)—R⁹]_(m) wherein SSrepresents a support material; R⁵, R⁶, and R⁹ are each independently anorganic group; R⁸ is an organic connecting group; n is 0 to 1; and m is1 to the resin capacity of the support material. Preferably, R⁹ includesa reactive site, which may optionally be protected by a protectinggroup. Preferably, R⁵ and R⁶ are independently a (C1–C14)alkyl group, a(C3–C14)cycloalkyl group, or a (C5–C12)aryl group), and R⁸ is a(C1–C1000)alkylene group. This material is a preferred example of anamine-modified-azlactone-functionalized support material having a linkerattached thereto, wherein NH—(R⁸)—NH is derived from a diamine.

Use of the functionalized support materials in solid phase synthesis(typically, solid phase organic synthesis) typically requires that thesupport material includes a linker with a reactive site at which one ormore reactions (e.g., synthetic organic reactions) can be conducted. Forexample, linker-functionalized supports can be used in buildingpolynucleotides and polypeptides, which can then be released from thelinker-functionalized supports. They can also be used in developingcombinatorial libraries by the systematic and repetitive covalentconnection of a set of different “building blocks” of varying structuresto the reactive site of the linker.

Thus, the support materials described above can be used as thefoundation on which such chemical reactions can be conducted if R⁷ andR⁹ include a reactive site. This reactive site can include a hydroxylgroup (e.g., wherein R⁷ is hydrogen) or an organic group capable ofbeing derivatized. Alternatively, the reactive site of R⁷ can beprotected with a protecting group, e.g., for a hydroxyl functionality,in cases where that is needed during the step of attaching a linkermolecule to the support material.

It should also be noted that the formulations of the support materialsdescribed above are used herein to refer to materials that include thefinal derivatized molecules prior to being removed from thelinker-functionalized support. In such cases R⁷ and R⁹ may not include areactive site; rather, they would include, for example, the desiredpolynucleotide or polypeptide or the desired set of molecules that formthe combinatorial library.

The present invention also provides methods that utilize such supportsas well as others. In one embodiment, the present invention provides amethod of solid phase synthesis that includes providing anazlactone-functionalized support; reacting the azlactone-functionalizedsupport with a linker molecule to form a linker-functionalized supporthaving a linker attached to the azlactone-functionalized support; andconducting one or more reactions on the linker functionalized support.Preferably, this latter step involves reacting the linker-functionalizedsupport with an organic molecule to form a covalent bond between thelinker and the organic molecule; and conducting one or more reactions onthe covalently bound organic molecule to produce a derivatized organicmolecule. The organic molecule is preferably a building block for acombinatorial library. Typically and preferably, the covalent bondformed between the linker and the organic molecule can be cleaved undermild conditions, such as, for example, the use of mild acids or bases,as is well known to those of skill in the art of solid phase synthesis.Thus, typically and preferably, the method involves cleaving thederivatized molecule from the linker-functionalized support at the siteof the covalent attachment to the linker.

Preferably, in the method described above, the linker-functionalizedsupport has the following formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein SS represents a support material;C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from an azlactone group,wherein R⁵ and R⁶ are each independently an organic group and n is 0 to1; NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents the linker, wherein R¹,R², R³, and R⁴ are each independently hydrogen or an organic group withthe proviso that at least one of R³ and R⁴ is an aromatic group, R⁷ ishydrogen, a protecting group (e.g., for an OH functional group), or anorganic group capable of being derivatized, and p is at least 1(preferably, 1 to 20, and more preferably, 1 to 2); and m is 1 to theresin capacity of the support material. In the methods using thismaterial, reactions occur at the —OR⁷ group.

In another preferred embodiment of the method described above, thelinker-functionalized support has the following formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(R⁸)—NH—C(O)—R⁹]_(m) wherein SSrepresents a support material; C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) isderived from an azlactone group, wherein R⁵ and R⁶ are eachindependently an organic group and n is 0 to 1; NH—(R⁸)—NH is derivedfrom a diamine, wherein R⁸ is an organic connecting group; C(O)—R⁹represents the linker, wherein R⁹ is an organic group; and m is 1 to theresin capacity of the support material. In the methods using thismaterial, reactions occur at the —R⁹ group.

Another preferred method of the present invention includes providing anamine-odified-azlactone-functionalized support; reacting theamine-modified-azlactone-functionalized support with a linker moleculeto form a linker-functionalized support having a linker attached to theamine-modified-azlactone-functionalized support; and conducting one ormore reactions on the linker-functionalized support. Preferably,conducting one or more reactions on the linker-functionalized supportincludes reacting it with an organic molecule to form a covalent bondbetween the linker and the organic molecule; and conducting one or morereactions on the covalently bound organic molecule to produce aderivatized organic molecule. Typically and preferably, the covalentbond formed between the linker and the organic molecule can be cleavedunder mild conditions, such as, for example, the use of mild acids orbases, as is well known to those of skill in the art of solid phasesynthesis. Thus, typically and preferably, the method involves cleavingthe derivatized molecule from the linker-functionalized support at thesite of the covalent attachment to the linker.

In yet another preferred embodiment, the present invention provides amethod of solid phase synthesis that includes providing alinker-functionalized support having the formulaSS—[NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m) wherein SS represents asupport material; NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents a linker,wherein R¹, R², R³, and R⁴ are each independently hydrogen or an organicgroup with the proviso that at least one of R³ and R⁴ is an aromaticgroup, R⁷ is hydrogen, a protecting group, or an organic group capableof being derivatized, and p is at least 1; and m is 1 to the resincapacity of the support material; and conducting one or more reactionson the linker-functionalized support. Preferably, this involves reactingthe linker-functionalized support with an organic molecule so as to forma covalent bond between the linker and the organic molecule; andconducting one or more reactions on the covalently bound organicmolecule to produce a derivatized organic molecule. Typically andpreferably, the covalent bond formed between the linker and the organicmolecule can be cleaved under mild conditions, such as those describedabove. Typically and preferably, the method involves cleaving thederivatized molecule from the linker-functionalized support.

Another preferred embodiment of the methods of the present inventionincludes providing an azlactone-functionalized support having a linkerattached thereto, which has the formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein SS represents a support material;C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from an azlactone group,wherein R⁵ and R⁶ are each independently an organic group and n is 0 to1; NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents the linker, wherein R¹,R², R³, and R⁴ are each independently hydrogen or an organic group withthe proviso that at least one of R³ and R⁴ is an aromatic group, R⁷ ishydrogen, a protecting group, or an organic group capable of beingderivatized, and p is at least 1; and m is 1 to the resin capacity ofthe support material; reacting the linker with an organic molecule toform a covalent bond between the linker and the organic molecule;conducting one or more reactions on the covalently bound organicmolecule to produce a derivatized organic molecule; and cleaving thederivatized molecule from the azlactone-functionalized support having alinker attached thereto.

Yet another preferred embodiment of the methods of the present inventionincludes providing a linker-functionalized support having the formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(R⁸)—NHC(O)—R⁹]_(m) wherein SSrepresents a support material; C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) isderived from an azlactone group, wherein R⁵ and R⁶ are eachindependently an organic group and n is 0 to 1; NH—(R⁸)—NH is derivedfrom a diamine, wherein R⁹ is an organic connecting group; C(O)—R⁹represents the linker, wherein R⁹ is an organic group; and m is 1 to theresin capacity of the support material; reacting the linker with anorganic molecule so as to form a covalent bond between the linker andthe organic molecule; conducting one or more reactions on the covalentlybound organic molecule to produce a derivatized organic molecule; andcleaving the derivatized molecule from the azlactone-functionalizedsupport having a linker attached thereto.

Whether directed to a method or a support, the present inventionincludes the following preferred embodiments he functionalized supportcan be in the form of a plurality of particles. Each R⁷ (or R⁹) can bethe same on any one particle, or the plurality of particles can includeat least two different R⁷ (or R⁹) groups. Alternatively, thefunctionalized support can be in the form of a membrane. Each R⁷ (or R⁹)can be the same on the membrane, or the membrane can include at leasttwo different R⁷ (or R⁹) groups.

DEFINITIONS

An “organic molecule” (i.e., the starting material) can be a monomer,oligomer, or polymer, although typically it is a monomer. These can beused as the “building blocks” in combinatorial chemistry. A “derivatizedorganic molecule” is an organic molecule that is different in some wayrelative to the starting organic molecule. The organic molecule andderivatized organic molecule can include heteroatoms and substituents asdescribed below with respect to the definition of “organic group.”Herein, an organic molecule can also include metals or metalloids, suchthat it could be classified as an organometallic molecule.

The term “organic group” means a hydrocarbon group (with optionalelements substituted for carbon and hydrogen, such as oxygen, nitrogen,sulfur, and silicon) that is classified as an aliphatic group, cyclicgroup, or combination of aliphatic and cyclic groups (e.g., alkaryl andaralkyl groups). In the context of the present invention, the organicgroups are those that do not interfere with chemical reactions thatoccur at the reactive site of the linker, such as occur in the formationof a derivatized organic molecule. The term “aliphatic group” means asaturated or unsaturated linear or branched hydrocarbon group. This termis used to encompass alkyl, alkenyl, and alkynyl groups, for example.The term “alkyl group” means a saturated linear or branched hydrocarbongroup including, for example, methyl, ethyl, isopropyl, t-butyl, heptyl,dodecyl, octadecyl, amyl, 2-ethylhexyl, and the like. The term “alkenylgroup” means an unsaturated, linear or branched hydrocarbon group withone or more carbon-carbon double bonds, such as a vinyl group. The term“alkynyl group” means an unsaturated, linear or branched hydrocarbongroup with one or more carbon-carbon triple bonds. The term “cyclicgroup” means a closed ring hydrocarbon group that is classified as analicyclic group, aromatic group, or heterocyclic group. The term“alicyclic group” means a cyclic hydrocarbon group having propertiesresembling those of aliphatic groups. The term “aromatic group” or “arylgroup” means a mono- or polynuclear aromatic hydrocarbon group. The term“heterocyclic group” means a closed ring hydrocarbon in which one ormore of the atoms in the ring is an element other than carbon (e.g.,nitrogen, oxygen, sulfur, etc.).

As a means of simplifying the discussion and recitation of certainterminology used throughout this application, the terms “group” and“moiety” are used to differentiate between chemical species that allowfor substitution or that may be substituted and those that do not allowor may not be so substituted. Thus, when the term “group” is used todescribe a chemical substituent, the described chemical materialincludes the unsubstituted group and that group with O, N, Si, or Satoms, for example, in the chain (as in an alkoxy group) as well ascarbonyl groups or other conventional substitution. Where the term“moiety” is used to describe a chemical compound or substituent, only anunsubstituted chemical material is intended to be included. For example,the phrase “alkyl group” is intended to include not only pure open chainsaturated hydrocarbon alkyl substituents, such as methyl, ethyl, propyl,t-butyl, and the like, but also alkyl substituents bearing furthersubstituents known in the art, such as hydroxy, alkoxy, alkylsulfonyl,halogen atoms, cyano, nitro, amino, carboxyl, etc. Thus, “alkyl group”includes ether groups, haloalkyls, nitroalkyls, carboxyalkyls,hydroxyalkyls, sulfoalkyls, etc. On the other hand, the phrase “alkylmoiety” is limited to the inclusion of only pure open chain saturatedhydrocarbon alkyl substituents, such as methyl, ethyl, propyl, t-butyl,and the like. A “hydrocarbyl moiety” refers to an organic moietycontaining only carbon and hydrogen atoms (no substituents orheteroatoms).

The term “organic connecting group” means an “organic group” which issituated between and joins at least two chemically reactive groups. Inthe case of the present invention this term is used preferably torepresent the “organic group” which joins two or more amino groups.

The term “linker molecule” refers to a molecule that can be permanentlyattached to a support material via covalent bonds to form a “linker”.The linker molecule (and linker) includes a reactive group capable ofbinding an organic molecule, which then can be derivitized and cleavedfrom the support material. Herein, linker molecule refers to the speciesprior to being attached to the support material and linker refers to thespecies after it has been attached to the support material.

The term “mild conditions” as it applies to cleavage of the covalentbond formed between the linker and the organic molecule refers toconditions that do not degrade, or otherwise affect, the derivatizedorganic molecule, but just removes it from the functionalized support.In general, these are conditions well known in the art of solid phasesynthesis.

The term “resin capacity” or “functional group density” means a measureof the amount of functionality of the support material (typically, inthe form of a resin), typically described in units such as moles/gram orequivalents/gram of resin.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The present invention provides functionalized supports and methods foruse for solid phase synthesis (typically, solid phase organicsynthesis), for example. These functionalized supports can be used forsynthesizing small molecules as well as large molecules (e.g.,biomolecules). Significantly, the functionalized support can be used insolid phase synthesis in which, for example, organic molecules (e.g.,monomers) are consecutively added to a chain or polymer, as occurs inthe formation of polypeptides, polynucleotides, etc., includingpeptidomimetics. The functionalized support can also be used to form acombinatorial library if desired, which is of significant interest in avariety of fields, particularly the pharmaceutical industry.

Support Material

Functionalized supports include a support material (often referred to asa base support or base polymer or base resin) and one or more functionalgroups, preferably, azlactone functional groups. The support materialcan be a pre-existing material to which functional groups, preferably,azlactone functional groups, are attached (e.g., through the use of highenergy radiation and free radical reactions), or the support materialand functionalization thereof can occur generally simultaneously (e.g.,through the use of free radical polymerization).

The support material can be a polymeric material that can be used inconventional solid phase synthesis. It is chosen such that it isgenerally insoluble in the solvents or other components used insynthetic reactions that occur during the course of solid phasesynthesis.

The support material can be organic or inorganic. It can be in the formof solids, gels, glasses, etc. It can be in the form of a plurality ofparticles (e.g., beads, pellets, or microspheres), fibers, a membrane(e.g., sheet or film), a disc, a ring, a tube, or a rod, for example.Preferably, it is in the form of a plurality of particles or a membrane.It can be swellable or nonswellable. It can be porous or nonporous. Itcan be pre-existing or made in situ (such that functionalization occursduring formation of the support material). Preferably, it is made insitu, as occurs in the formation of vinylazlactone/methylenebisacrylamide copolymer beads.

Examples of useable pre-existing support materials are described in G.B. Fields et al., Int. J. Peptide Protein Res., 35, 161 (1990) and G. B.Fields et al., in Synthetic Peptides: A User's Guide, G. A. Grant, Ed.,pages 77–183, W. H. Freeman and Co., New York, N.Y. (1992). Preferably,the support material is in the form of an organic polymeric material,such as polystyrenes, polyalkylenes, nylons, polysulfones,polyacrylates, polycarbonates, polyesters, polyimides, polyurethanes,etc. For pre-existing support materials, a preferred support material ispolystyrene. Included in the term “polystyrene” are polymers that havebeen substituted to some extent with substituents that are not capableof reaction under the conditions generally used for solid phasesynthesis of biomolecules, e.g., substituents such as alkyl and alkoxygroups. In order to increase the stability and insolubility in organicsolvents, polystyrene resins are typically crosslinked with, forexample, divinyl benzene or butadiene.

Functionalized Supports

Preferably, the support material includes functional groups to whichlinker molecules can be attached for building large or small organiccompounds. Suitable functional groups include electrophilic groups suchas epoxide or oxirane groups, N-hydroxysuccinimide ester groups,sulfonyl ester groups, iodoacetyl groups, aldehyde groups,imidazolylcarbamate groups, chlorotriazine groups, or other groupscapable of reacting to form covalent bonds with linker molecules,particularly those linker molecules containing amino groups.

In one preferred embodiment, the functional groups are azlactone groups.

Azlactone-functionalized supports have been described in U.S. Pat. No.5,403,902 (Heilmann et al.), for example. They are described as beinguseful reactive supports for the attachment of functional materials toprovide adduct beads. The adduct beads are useful as complexing agents,catalysts, polymeric reagents, chromatographic supports, and as enzyme-or other biomacromolecule-bearing supports. Azlactone beads have highbinding capacity with functional materials even when the beads arehighly crosslinked and swell very modestly, e.g., threefold or less, inwater.

It has now been found that azlactone-functionalized supports, such asthese, can be reacted with a linker molecule to form a linker which canbe further reacted with a building block molecule (i.e., organicmolecules typically used in combinatorial chemistry to build largermolecules such as polypeptides and polynucleotides) through a covalentbond. With certain linkers the covalent bond is cleavable under mildconditions. The building block molecule can then be subjected tonumerous chemical reactions using, for example, a combinatorialsynthetic scheme to produce a library of compounds attached to thesupport via the linker. When the covalent bond between the linker andthe covalently bound building block molecule is cleaved under mildconditions, the library of organic compounds is released, regeneratingthe active support (i.e., the functionalized support with linkersattached thereto).

Particularly preferred azlactone-functionalized supports include vinylazlactone copolymers, such as those described in U.S. Pat. No. 5,403,902(Heilman et al.). Most preferred azlactone-functionalized supports arevinyl azlactone/methylenebisacrylamide copolymers, such as thosecommercially available under the trade designation EMPHAZE AB 1 fromMinnesota Mining and Manufacturing Company (St. Paul, Minn.) orULTRALINK Biosupport Medium from Pierce Scientific (Rockford, Ill.).These copolymers are extremely stable to strongly acidic and basicconditions, and thus are ideal base supports for solid phase synthesis.

Typically and preferably, linker molecules can be added to suchfunctionalized supports to create reactive sites for solid phasesynthesis. The term “linker molecule” refers to a molecule that can bepermanently attached to a support material via covalent bonds to form a“linker”. The linker molecule (and linker) includes a reactive groupcapable of binding an organic molecule, which then can be derivitizedand cleaved from the support material, if desired.

The linker is preferably chemically stable to the reaction conditionsnecessary to derivatize the organic molecule (e.g., and build acombinatorial library). It also preferably is chosen to allow thesynthesized molecules to be easily cleaved from the support. The linkermay include a protecting group (e.g., a hydroxylprotecting group) at thereactive site, if desired, which can be removed prior to conducting thedesired chemical reactions for building larger molecules, for example.

Preferred linker molecules include, but are not limited to,aminoalcohols having the structure H₂N—(C(R¹)(R²))_(p)—C(R³)(R⁴)—OH,such as 2-amino-1-phenylethanol, 2-amino-1-(4-methoxyphenyl)ethanol,2-amino-1-methyl-1-phenylethanol, 2-amino-1,1-diphenylethanol,3-amino-1-phenylpropanol, 2-amino-1-phenylpropanol, and the like. Suchmolecules are readily prepared by cyanosilylation/reduction of aldehydesand ketones as described in Evans et al., J. Org. Chem., 39, 914 (1974)and in U.S. Pat. No. 4,918,231 (Krepski et al.). These linker moleculesprovide benzylic alcohol functionality similar to the familiar Wang andRink linkers (described, for example, in Wang, J. Amer. Chem. Soc., 95,1328 (1973) and Rink, Tetrahedron Letters, 28, 3787 (1987)) commonlyused in solid phase synthesis, but in addition contain aminefunctionality useful for providing stable amide bonds to the supportmaterial.

Optically active amino alcohols are other examples of linker molecules.They offer the possibility of conducting asymmetric synthetictransformations on the attached organic molecule. Specific, well-knownexamples include erythro-alpha-(1-aminoethyl)benzyl alcohol (also knownas (1S,2R)-(+)-norephedrine), (R)-(−)-norepinephrine,(S)-(+)-norepinephrine, L-erythro-2-(methylamino)-1-phenylpropanol (akal-ephedrine), D-threo-2-(methylamino)-1-phenylpropanol (also known asd-pseudoephedrine), and d-2-amino-1-phenylethanol.

In addition to the preferred linker molecules described above, many ofthe traditional linker molecules commonly utilized for solid phasesynthesis can also be used with azlactone-functionalized supportsprovided that these supports are suitably derivatized to allowattachment of the traditional linker molecules. Preferably, thisderivatization process involves reaction of the azlactone group with anexcess of a polyamine, to produce anamine-modified-azlactone-functionalized support. Examples of polyaminesinclude primary polyamines, such as ethylenediamine, 1,3-propanediamine,1,3-diamino-2-hydroxypropane, 1,6-hexanediamine,tris-(2-aminoethyl)amine, and the like; and polyetherpolyamines, such as4,7,10-trioxa-1,13-tridecanediamine, 3,6-dioxa-1,8-diaminooctane,amine-terminated polyethyleneglycol and polypropyleneglycol homopolymersand copolymers; and the like. Preferably, the polyamines are diamines,such as ethylenediamine, 1,3-propanediamine,1,3-diamino-2-hydroxypropane, or 1,6-hexanediamine. In a second step,carboxyl functional linker molecules can be reacted with the amine toform an amide bond to the support. Examples of suitable linker moleculesinclude, but are not limited to, 4-hydroxymethylbenzoic acid,4-hydroxymethylphenoxyacetic acid,4-hydroxymethyl-3-methoxyphenoxybutyric acid,4-hydroxymethylphenylacetic acid, 4-bromoacetylphenoxyacetic acid,4-(diphenylhydroxymethyl)benzoic acid,4-hydroxymethyl-2-methoxy-5-nitrophenoxybutyric acid, phenoxyacetic acidand phenoxybutyric acid analogs of Rink acid and Rink amide linkermolecules and Sieber amide linker molecules (described, for example, inRink, Tetrahedron Letters, 28, 3787 (1987) and Sieber, TetrahedronLetters, 28, 2107 (1987)), 4-sulfamylbenzoic acid, 4-sulfamylbutyricacid, 4-formylphenoxyacetic acid, 4-(4-formyl-3-methoxyphenoxy)butyricacid, 4-formyl-3,5-dimethoxyphenoxyacetic acid, 3-formylindol-1-ylaceticacid, and the like. This synthetic scheme preferably results in a resinof the general structureSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH-(organic group fromamine)-NH—C(O)-linker]_(m).

The linker molecules can be attached to the support material usingconventional attachment chemistry, such as carbodiimide chemistry, mixedanhydride chemistry, and the like. Such techniques are well known to oneof skill in the art.

Once attached to the functionalized support, the linker provides one ormore reactive sites for subsequent reaction, such as those that occur insolid phase synthesis (typically, organic synthesis). Suchfunctionalized supports having a linker attached thereto are referred toherein as linker-functionalized supports. The reactive site can includea hydroxyl group or an organic group capable of being derivatized.Alternatively, the reactive site can be protected by a protecting group,e.g., for a hydroxyl functionality, in cases where that is needed duringthe step of attaching a linker molecule to the support material.Examples of such protecting groups include t-butyldimethylsilyl,triphenylmethyl, and others well known in the art (see, for example,Harrison and Harrison, Compendium of Organic Synthetic Methods, pages124–131, John Wiley and Sons, New York, 1971). Such protecting groupsare removed during the process of conducting reactions on the linker.

It should also be noted that the formulations of the support materialsdescribed herein are used herein to refer to materials that include thefinal derivatized molecules prior to being removed from thelinker-functionalized support. In such cases the linkers (which include,for example R⁷ and R⁹ in the formulations herein) may not include areactive site; rather, they would include, for example, the desiredpolynucleotide or polypeptide or the desired set of molecules that formthe combinatorial library. As a result they can be quite large.

Preferably, a linker-functionalized support has the following formulaSS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein SS represents a support material; R¹, R², R³, and R⁴ are eachindependently hydrogen or an organic group (preferably, having up toabout 20 carbon atoms) with the proviso that at least one of R³ and R⁴is an aromatic group; R⁷ is hydrogen or an organic group; R⁵ and R⁶ areeach independently an organic group (preferably, having up to about 20carbon atoms); n is 0 to 1; p is at least 1 (preferably, 1 to 20, andmore preferably, 1 to 2); and m is 1 to the resin capacity of thesupport material. Preferably, C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) isderived from an azlactone group and NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)represents the linker. Preferably, when OR⁷ is the attachment site foran organic molecule, R⁷ is hydrogen or an organic group capable of beingderivatized, as in combinatorial chemistry, for example. Alternatively,the organic group could be a protecting group for, e.g., a hydroxylfunctionality, in cases where that is needed during the step ofattaching the linker to the support. Prior to any reactions beingconducted on the linker-functionalized support, the protecting group isremoved. For certain embodiments, R⁷ can include the final desiredproduct. As a result it can be quite large, including polynucleotidesand polypeptides, for example.

Another preferred linker-functionalized support has the followingformula SS—[C(O)NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(R⁸)—NH—C(O)—R⁹]_(m)wherein SS represents a support material; R⁵ and R⁶ are eachindependently an organic group (preferably, having up to about 20 carbonatoms); R⁹ is an organic group; R⁸ is an organic connecting group(preferably, having up to about 1000 carbon atoms); n is 0 to 1; and mis 1 to the resin capacity of the support material. R⁸ can be any linearor branched organic group and is preferably derived from diamines.Examples of such diamines include primary diamines, such asethylenediamine, 1,3-propanediamine, 1,3-diamino-2-hydroxypropane and1,6-hexanediamine, and the like; and polyetherdiamines, such as3,6-dioxa-1,8-diaminooctane, amine-terminated polyethyleneglycol andpolypropyleneglycol homopolymers and copolymers; and the like.Preferably, C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from anazlactone group, NH—(R⁸)—NH is derived from a diamine, and C(O)—R⁹represents the linker. Preferably, NH—(R⁸)—NH is derived fromethylenediamine, 1,3-propanediamine, 1,3-diamino-2-hydroxypropane, or1,6-hexanediamine.

Preferably, when C(O)—R⁹ is the linker, it includes an attachment sitefor an organic molecule. Such attachment sites can be the same as thosedescribed above for R⁷ (e.g., hydroxyl group, an organic group capableof being derivatized, or a protecting group). Preferably, C(O)—R⁹ isderived from 4-hydroxymethylbenzoic acid, 4-hydroxymethylphenoxyaceticacid, 4-hydroxymethyl-3-methoxyphenoxybutyric acid,4-hydroxymethylphenylacetic acid, 4-bromoacetylphenoxyacetic acid,4-(diphenylhydroxymethyl)benzoic acid,4-hydroxymethyl-2-methoxy-5-nitrophenoxybutyric acid, phenoxyacetic acidand phenoxybutyric acid analogs of Rink acid and Rink amide linkermolecules and Sieber amide linker molecules, 4-sulfamylbenzoic acid,4-sulfamylbutyric acid, 4-formylphenoxyacetic acid,4-(4-formyl-3-methoxyphenoxy)butyric acid,4-formyl-3,5-dimethoxyphenoxyacetic acid, or 3-formylindol-1-ylaceticacid. For certain embodiments, R⁹ can include the final desired product.As a result it can be quite large, including polynucleotides andpolypeptides, for example.

Alternatively, a linker-functionalized support does not necessarily haveto be derived from azlactone functionality, but can have the followingformula SS—[NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m) wherein SS representsa support material; R¹, R², R³, and R⁴ are each independently hydrogenor an organic group with the proviso that at least one of R³ and R⁴ isan aromatic group; R⁷ is hydrogen or an organic group; p is at least 1(preferably, 1 to 20, and more preferably, 1 to 2); and m is 1 to theresin capacity of the support material. Preferably,NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents a linker. It is typicallyattached to the support material through a carbonyl group, therebyforming an amide linkage. Preferably, when OR⁷ is the attachment sitefor an organic molecule, R⁷ is hydrogen, an organic group capable ofbeing derivatized, or a protecting group, which would be removed priorto any reactions being conducted on the linker-functionalized support.More preferably, R⁷ is hydrogen. For certain embodiments, R⁷ can includethe final desired product. As a result it can be quite large, includingpolynucleotides and polypeptides, for example.

When the functionalized supports include organic groups at the R¹, R²,R³, R⁴, R⁵, R⁶ and R⁸ positions, these can be of any size orfunctionality that do not interfere with the solid phase synthesisreactions. Preferably, R¹, R², R³, R⁴, R⁵, and R⁶ are each independentlyalkyl groups (preferably, containing 1 to 14 carbon atoms, and morepreferably (C1–C14)alkyl moieties) or cycloalkyl groups (preferably,containing 3 to 14 carbon atoms, and more preferably (C3–C14)cycloalkylmoieties), aryl groups (preferably, containing 5 to 12 ring atoms, andmore preferably (C5–C12)aryl moieties). Preferably, at least one of R³and R⁴ is an aryl group (preferably, containing 5 to 12 ring atoms). Anytwo of the groups R¹ and R² or R⁵ and R⁶ taken together with the carbonto which they are joined can form a carbocyclic ring, preferablycontaining 4 to 12 ring atoms. Preferably, R⁸ is an alkylene group (morepreferably, an alkylene moiety) having up to about 1000 carbon atoms.

Functionalized supports (including linker-functionalized supports) ofthe present invention can have the same or mixtures of functional groupsand/or linkers. For example, support materials as described herein caninclude at least two different R⁷ (or R⁹) groups. If particles are used,this can result from blending two different samples of particles, eachwith a different R⁷ (or R⁹) group. Alternatively, a membrane can includeat least two different R⁷ (or R⁹) groups.

Base functionalized supports can be prepared by methods well known inthe art, and many are available commercially (e.g., from variouscompanies such as Novabiochem, BiORad, Pierce, Amersham-Pharmacia, RappPolymere, Polymer Laboratories, Sigma-Aldrich, Millipore, EMSeparations, etc.). Methods for the preparation ofazlactone-functionalized supports are described, for example, in U.S.Pat. No. 5,403,902 (Heilmann et al.). This patent describes thepreparation of particulate supports by suspension or dispersionpolymerization processes. Other methods for preparing usefulazlactone-functionalized supports are described in U.S. Pat. No.5,262,484 (Coleman et al.), which describes graft copolymers andarticles prepared therefrom, U.S. Pat. No. 5,292,514 (Capecchi et al.),which describes functionalized substrates, U.S. Pat. No. 5,451,453(Gagnon et al.), which describes porous supports, U.S. Pat. No.5,486,358 (Coleman et al.), which describes polymer blends and articlesprepared therefrom, U.S. Pat. No. 5,510,421 (Dennison et al.), whichdescribes membrane supports, and U.S. Pat. No. 5,993,935 (Rasmussen etal.), which describes bead/porous matrix composites.

Use of the Functionalized Supports

The functionalized supports are preferably used to covalently attach alinker molecule and to provide a starting point for solid phasesynthesis of a compound, which may or may not be polymeric. For example,in creating a combinatorial library, the functional groups with linkerand organic molecule attached thereto can be divided into groups andthen chemically modified by introduction of substituents to form aseries of analogs. Alternatively, conventional formation of a polymer(e.g., homopolymer, copolymer, terpolymer, etc.) by stepwise addition ofmonomers can occur.

Conventional solid phase synthetic techniques can be used. Suchsynthetic techniques can include the use of protecting groups. These canbe deprotected using appropriate cleavage reagents well known to thoseof skill in the art.

After synthesis is complete, cleavage conditions are used to remove themodified organic molecule (i.e., organic compound), preferably bycleaving the covalent bond between the linker and the organic molecule.Preferably, the cleavage conditions are mild, whether they be acidic orbasic. Typically, mild conditions involve the use of acids (particularlyacids having an H₀ of −5 or higher, as defined by J. P. Tam et al. inThe Peptides, Vol. 9, S. Udenfreind and J. Eienhofer, Eds., pages185–248, Academic Press, New York, N.Y. (1987)), such as hydrochloric,acetic, sulfuric, and trifluoroacetic acid. Preferably, trifluoroaceticacid is used. Basic cleavage conditions may also be used through the useof, for example, sodium hydroxide or ammonia solutions. Other usefulcommon methods of cleavage are reviewed in numerous literature articles,for example, in “The Combinatorial Chemistry Catalog” published annuallyby Calbiochem-Novabiochem, San Diego, Calif.

The method of using the functionalized supports described herein isparticularly useful in preparing a combinatorial library. Specifically,in making such a library, a plurality of reaction vessels are provided,each containing a functionalized support with a linker attached thereto.A different monomer, each capable of reacting with the linker on thefunctionalized support, is provided in each vessel. Additional monomersare coupled to the growing oligomer chain, with the identity and orderof monomers documented to enable synthesis of a plurality ofsupport-bound, chemically distinct oligomers. This last step may involvea “split/mix” approach, wherein after every monomer addition, thecontents of the reaction vessels are alternatively divided and mixed ina way that provides for a completely diverse set of ligands. Thedistinct oligomers in the combinatorial library so provided are thenscreened for activity, generally by screening individual sublibrariescontaining mixtures of distinct oligomers, identifying activesublibraries, and then determining the oligomeric compounds of interestby generating different sublibraries and cross-correlating the resultsobtained.

EXAMPLES

The following examples are given to illustrate, but not limit, the scopeof this invention. Unless otherwise indicated, all parts and percentagesare by weight and all molecular weights are weight average molecularweights.

Example 1

Coupling of 2-amino-1-phenylethanol (Aldrich Chemical Co., Milwaukee,Wis.) to EMPHAZE AB 1 beads (Minnesota Mining and Manufacturing Company)1 molar (1M) solutions were prepared of the aminoalcohol in (a)dimethylformamide/deionized water (12 mL/4 mL) and (b)dimethylsulfoxide/deionized water (12 mL/4 mL). To each solution wasadded 1.0 gram (g) AB 1 beads, and each mixture was tumbled for 3.5hours (hrs). Workup was accomplished by filtering, washing thederivatized beads with acetone (3 times), deionized water, 0.1 normal(0.1N) HCl (2 times), then deionized water until the filtrate wasneutral to pH paper. Evaluation by a cation exchange procedure forlysozyme, as described in U.S. Pat. No. 5,561,097 (Gleason et al.),indicated a 70% coupling efficiency of the linker in both reactions.

Example 2

Procedures similar to those of Example 1 were used to couple2-amino-1-phenylethanol to 140 micrometer (μ) diameterazlactone-functional beads. Details are listed in Table 1. Lysozymecation exchange testing was used to estimate coupling efficiency of thelinker.

TABLE 1 DMSO/water (v/v) Time (hrs) EEDQ¹ Coupling Efficiency (%) a) 4:11 + 33 b) 4:1 3 − 29 c) 1:4 2 + 62 d) 1:4 2 − 70 e) 1:4 3 − 78 ¹Reactiondone in the presence (+) or absence (−) of 0.1 M2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline.

Example 3

Attachment and release of benzoic acid to modified beads wasaccomplished using the following methods: benzoic acid was coupled tothe beads of Example 2d by the procedure of Valerio et al., Int. J.Peptide Res., 44, 158–165 (1994) using diisopropylcarbodiimide and4-dimethylaminopyridine in 25/75:volume/volume (25%) DMF/CH₂Cl₂.Specifically, 2 milliliters (mL) of wet beads were placed in a 15 mLpolypropylene disposable chromatography column and mixed with 7 mL of 1Nsodium hydroxide for 1 hour. The sodium hydroxide solution was drainedoff and the beads were washed 3 times with 10 mL of deionized water, 3times with 10 mL of acetone, then 2 times with 10 mL of 25% DMF/CH₂Cl₂mixture. The damp beads were then mixed with a solution of 37 milligrams(mg) benzoic acid, 47 microliters (μL) diisopropyl-carbodiimide, and 4mg 4-dimethylaminopyridine in 3 mL 25% DMF/CH₂Cl₂ mixture. The mix wasallowed to react overnight at room temperature, filtered, and washedwith 10 mL 25% DMF/CH₂Cl₂ mixture, 3 times with 10 mL acetone, 3 timeswith 10 mL ethanol, and 3 times with deionized water. The derivatizedbeads were mixed with 8 mL of 0.1N sodium hydroxide for 1 hour and thesodium hydroxide extract was drained off. Second and third 8 mLhydrolysis extracts were made, using 1.0N and 2.0N sodium hydroxide,respectively. Anion exchange-SR extraction disks commercially availableunder the trade designation EMPORE from Minnesota Mining andManufacturing Company were preconditioned according to themanufacturer's recommendations, then a hydrolysis extract was passedthrough the membrane using aspirator vacuum, and the membrane was washed2 times with deionized water. The filtration apparatus was transferredto a clean filter flask, and the membrane was eluted 2 times with 10 mLconcentrated ammonia. The eluate was finally evaporated to dryness undervacuum, leaving a small amount of white residue. From the 2.0N extract,the ammonia eluate was acidified to pH 1 with concentrated hydrochloricacid, then passed through a preconditioned EMPORE C18 solid phaseextraction disk. The disk was then allowed to dry for 1 hour and eluted2 times with 10 mL acetonitrile. GC-MS analysis of the eluate residueupon evaporation indicated the presence of benzoic acid as a majorcomponent. The other extracts also contained benzoic acid. Extractiondisks commercially available under the trade designation EMPORE C8 fromMinnesota Mining and Manufacturing Co. were also useful for recoveringthe hydrolysis products.

Example 4

Benzoic acid was coupled to 1 mL of the beads of Example 2e by theprocedure described in Example 3. After coupling and washing, the beadswere mixed with 7 mL concentrated ammonia for 1 hour. The ammoniasolution was drained off, and the ammonia hydrolysis procedure wasrepeated a second time. The combined ammonia solutions were evaporatedunder vacuum to give a white residue. GC-MS of this residue identifiedbenzamide as the major component.

Examples 3 and 4 illustrate that the linker of Example 1 can be used toattach and subsequently release an appropriate organic molecule undermild basic hydrolysis conditions.

Example 5

2-Amino-1-(4-methoxyphenyl)ethanol was prepared according to theprocedure of Evans et al., J. Org. Chem., 39, 914 (1974) bycyanosilylation of 4-methoxybenzaldehyde followed by lithium aluminumhydride reduction. The crude aminoalcohol (36.9 g) was dissolved in 150mL of hot ethanol on a steam bath. To this mixture was slowly added12.83 g of fumaric acid. The precipitated salt was filtered and washedwith additional ethanol. Recrystallization from methanol providedgreater than 99% pure 2:1 amine:fumarate salt.

EMPHAZE AB 1 beads (250 mg) and the above fumarate salt (740 mg) in 4.5mL deionized water were allowed to react for 2 hours. The derivatizedbeads were filtered, washed with DMSO (2 times), acetone (2 times),deionized water, 0.1N HCl, then deionized water until the filtrate wasof neutral pH. Lysozyme cation exchange analysis indicated an 82%coupling efficiency. The derizatized beads were treated 3 times insuccession, 1 hour each, with 4 mL volumes of 5% trifluoroacetic acid(TFA) in CH₂Cl₂. The beads were washed with deionized water (3 times),acetone (3 times), and 25% DMF/CH₂Cl₂. Benzoic acid was coupled to thesebeads using a procedure similar to that of Example 3. Benzoic acid couldbe released from these beads using low concentrations of TFA (1%, 2%,5%) in CH₂Cl₂.

Example 6

EMPHAZE AB 1 beads (25 g) were derivatized by reaction with 300 mL of 1Methylenediamine in deionized water for 2 hours at room temperature. Thederivatized beads were washed with deionized water (2×), 0.1N HCl (2×),0.0001N HCl, and stored in 20% ethanol/water until needed. Titrationindicated the amine content to be 42 μmol of amine per milliliter ofbeads.

3-Formylindol-1-ylacetic acid was prepared and coupled to the beadsabove according to the process of EP 0 801 083 A2 (Estep et al.) usingdiisopropyl carbodiimide, N,N-diisopropylethylamine, andN-hydroxybenzotriazole in DMF/CH₂Cl₂. Benzylamine was reductivelycoupled to this bead-linker using sodium cyanoborohydride in 0.5Macetate buffer, pH 5, by the procedure in the same document. The productwas then acetylated using acetic anhydride/triethylamine. The acetylatedamine was released from the resin by treatment with 50% TFA/CH₂Cl₂ for 4hours. The filtrate was evaporated to dryness and the residue evaluatedby NMR and mass spectroscopy to show that the major product wasN-benzylacetamide. This example demonstrates the feasibility ofconducting solid phase synthesis using azlactone-functionalized beads.

The complete disclosures of the patents, patent documents, andpublications cited herein are incorporated by reference in theirentirety as if each were individually incorporated. Variousmodifications and alterations to this invention will become apparent tothose skilled in the art without departing from the scope and spirit ofthis invention. It should be understood that this invention is notintended to be unduly limited by the illustrative embodiments andexamples set forth herein and that such examples and embodiments arepresented by way of example only with the scope of the inventionintended to be limited only by the claims set forth herein as follows.

1. A method of solid phase synthesis, the method comprising: providingan azlactone-functionalized support; reacting an azlactone group of theazlactone-functionalized support with a linker molecule to form alinker-functionalized support having a covalently attached linker group;and reacting the linker group of the linker-functionalized support withan organic molecule to form a covalent bond between the linker and theorganic molecule; and conducting one or more reactions on the covalentlybound organic molecule to produce a derivatized organic molecule.
 2. Themethod of claim 1 wherein the covalent bond formed between the linkerand the organic molecule can be cleaved under mild conditions.
 3. Themethod of claim 2 wherein mild conditions comprise mild acidic or mildbasic conditions.
 4. The method of claim 1 further comprising cleavingthe derivatized molecule from the linker-functionalized support.
 5. Themethod of claim 1 wherein the organic molecule is a building block for acombinatorial library.
 6. The method of claim 1 wherein the derivatizedorganic molecule is a polypeptide or polynucleotide.
 7. The method ofclaim 1 wherein the linker-functionalized support has the followingformula:SS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein: SS represents a support material;C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from an azlactone group,wherein R⁵ and R⁶ are each independently an organic group and n is 0 to1; NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents the linker, wherein R¹,R², R³, and R⁴ are each independently hydrogen or an organic group withthe proviso that at least one of R³ and R⁴ is an aromatic group, R⁷ ishydrogen, a protecting group, or an organic group capable of beingderivatized, and p is at least 1; and m is 1 to the resin capacity ofthe support material; and further wherein reacting thelinker-functionalized support with an organic molecule occurs at the—OR⁷ group.
 8. The method of claim 7 wherein p is 1 to
 20. 9. The methodof claim 7 wherein R⁷ is hydrogen.
 10. The method of claim 7 wherein R⁷is a protecting group and conducting one or more reactions on the linkerattached to the azlactone-functionalized support comprises removing theprotecting group.
 11. The method of claim 1 wherein theazlactone-functionalized support is in the form of a plurality ofparticles or a membrane.
 12. A method of solid phase synthesis, themethod comprising: providing an amine-modified-azlactone-functionalizedsupport; reacting the amine-modified-azlactone-functionalized supportwith a linker molecule to form a linker-functionalized support having acovalently attached linker; and conducting one or more reactions on thelinker-functionalized support.
 13. The method of claim 12 whereinconducting one or more reactions on the linker-functionalized supportcomprises: reacting the linker-functionalized support with an organicmolecule to form a covalent bond between the linker and the organicmolecule; and conducting one or more reactions on the covalently boundorganic molecule to produce a derivatized organic molecule.
 14. Themethod of claim 13 wherein the covalent bond formed between the linkerand the organic molecule can be cleaved under mild conditions.
 15. Themethod of claim 14 wherein mild conditions comprise mild acidic or mildbasic conditions.
 16. The method of claim 13 further comprising cleavingthe derivatized molecule from the linker-functionalized support.
 17. Themethod of claim 12 wherein the linker-functionalized support has thefollowing formula:SS—[C(O)—NH—C(R⁵)(R⁶)—(CH²)_(n)—C(O)—NH—(R⁸)—NH—C(O)—R⁹]_(m) wherein: SSrepresents a support material; C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) isderived from an azlactone group, wherein R⁵ and R⁶ are eachindependently an organic group and n is 0 to 1; NH—(R⁸)—NH is derivedfrom a diamine, wherein R⁸ is an organic connecting group; C(O)—R⁹represents the linker, wherein R⁹ is an organic group; and m is 1 to theresin capacity of the support material; and further wherein reacting thelinker-functionalized support with an organic molecule occurs at the —R⁹group.
 18. A method of solid phase synthesis, the method comprising:providing a linker-functionalized support having a covalently attachedlinker, the linker-functionalized support having a formula:SS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein: SS represents a support material;C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from an azlactone group,wherein R⁵ and R⁶ are each independently an organic group and n is 0 to1; NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents a linker, wherein R¹,R², R³, and R⁴ are each independently hydrogen or an organic group withthe proviso that at least one of R³ and R⁴ is an aromatic group, R⁷ ishydrogen, a protecting group, or an organic group capable of beingderivatized, and p is at least 1; and m is 1 to the resin capacity ofthe support material; and conducting one or more reactions on thelinker-functionalized support.
 19. A method of solid phase synthesis,the method comprising: providing a linker-functionalized support havinga covalently attached linker, the linker-functionalized support formula:SS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R⁸)—NH—C(O)—R⁹]_(m) wherein:SS represents a support material; C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) isderived from an azlactone group, wherein R⁵ and R⁶ are eachindependently an organic group and n is 0 to 1; NH—(R⁸)—NH is derivedfrom a diamine, wherein R⁸ is an organic connecting group; C(O)—R⁹represents the linker, wherein R⁹ is an organic group; and m is 1 to theresin capacity of the support material; and conducting one or morereactions on the linker-functionalized support.
 20. A method of solidphase synthesis, the method comprising: providing anazlactone-functionalized support; reacting the azlactone-functionalizedsupport with a linker molecule to form a linker-functionalized supporthaving a linker attached thereto, which has the formula:SS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷)]_(m)wherein: SS represents a support material;C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) is derived from an azlactone group,wherein R⁵ and R⁶ are each independently an organic group and n is 0 to1; NH—(C(R¹)(R²))_(p)—C(R³)(R⁴)(OR⁷) represents a linker, wherein R¹,R², R³, and R⁴ are each independently hydrogen or an organic group withthe proviso that at least one of R³ and R⁴ is an aromatic group, R⁷ ishydrogen, a protecting group, or an organic group capable of beingderivatized, and p is at least 1; and m is 1 to the resin capacity ofthe support material; reacting the linker with an organic molecule toform a covalent bond between the linker and the organic molecule;conducting one or more reactions on the covalently bound organicmolecule to produce a derivatized organic molecule; and cleaving thederivatized molecule from the linker-functionalized support.
 21. Amethod of solid phase synthesis, the method comprising: providing anazlactone-functionalized support; reacting the azlactone-functionalizedsupport with a diamine to forma anamine-modified-azlactone-functionalized support; reacting theamine-modified-azlactone-functionalized support with a linker moleculeto form a linker-functionalized support having a covalently attachedlinker, the linker-functionalized support having the formula:SS—[C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O)—NH—(C(R⁸)—NHC(O)—R⁹]_(m) wherein:SS represents a support material; C(O)—NH—C(R⁵)(R⁶)—(CH₂)_(n)—C(O) isderived from an azlactone group, wherein R⁵ and R⁶ are eachindependently an organic group and n is 0 to 1; NH—(R⁸)—NH is derivedfrom the diamine, wherein R⁸ is an organic connecting group; C(O)—R⁹represents the linker, wherein R⁹ is an organic group; and m is 1 to theresin capacity of the support material; reacting the linker with anorganic molecule so as to form a covalent bond between the linker andthe organic molecule; conducting one or more reactions on the covalentlybound organic molecule to produce a derivatized organic molecule; andcleaving the derivatized molecule from the linker-functionalizedsupport.
 22. The method of claim 21 wherein C(O)—R⁹ is derived from4-hydroxymethylbenzoic acid, 4hydroxymethylphenoxyacetic acid,4-hydroxymethyl-3-methoxyphenoxybutyric acid,4-hydroxymethylphenylacetic acid, 4-bromoacetylphenoxyacetic acid,4-(diphenylhydroxymethyl)benzoic acid,4-hydroxymethyl-2-methoxy-5-nitrophenoxybutyric acid, phenoxyacetic acidand phenoxybutyric acid analogs of Rink acid and Rink amide linkermolecules and Sieber amide linker molecules, 4sulfamylbenzoic acid,4-sulfamylbutyric acid, 4-formylphenoxyacetic acid,4-(4-formyl-3-methoxyphenoxy)butyric acid,4-formyl-3,5-dimethoxyphenoxyacetic acid, or 3-formylindol-1-ylaceticacid.
 23. The method of claim 21 wherein NH—(R⁸)—NH is derived fromethylenediamine, 1,3-propanediamine, 1,3-diamino-2-hydroxypropane, or1,6-hexanediamine.